CN112390848A - A Margarita powder polypeptide extract with antioxidant effect, and its preparation method - Google Patents
A Margarita powder polypeptide extract with antioxidant effect, and its preparation method Download PDFInfo
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- 238000000034 method Methods 0.000 claims abstract description 32
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- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
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- BJRNKVDFDLYUGJ-UHFFFAOYSA-N p-hydroxyphenyl beta-D-alloside Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=C(O)C=C1 BJRNKVDFDLYUGJ-UHFFFAOYSA-N 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/145—Extraction; Separation; Purification by extraction or solubilisation
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
- A61K8/987—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of species other than mammals or birds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/84—Products or compounds obtained by lyophilisation, freeze-drying
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- Animal Behavior & Ethology (AREA)
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- Medicinal Chemistry (AREA)
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- Proteomics, Peptides & Aminoacids (AREA)
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Abstract
The invention provides a pearl powder polypeptide extract with an antioxidation effect and a preparation method thereof. According to the invention, the pearl powder water-soluble protein is extracted by a weak acid method, reaction conditions (pH, temperature, reaction time and the like) are optimized, the hydroxyl radical scavenging effect of a sample is analyzed, the protein content conditions with different molecular weights are measured, and tests prove that the extract has a good skin whitening effect. The invention adopts normal-temperature acetic acid solution for auxiliary extraction to obtain a matrix protein extract with high water-soluble protein purity (more than or equal to 40.05 percent), the obtained proteins are all below 10,000Da, the polypeptide content of 0-2000 Da accounts for more than 70 percent of the total protein, the absorption by a human body is facilitated, and the matrix protein extract has obvious hydroxyl free radical removing effect in vitro; and the method has the advantages of simple process, no fine separation step, high preparation efficiency and low cost, and is suitable for subsequent pilot plant test and industrial production.
Description
(I) technical field
The invention relates to a pearl powder polypeptide extract with an antioxidant effect and a preparation method thereof.
(II) background of the invention
The pearl has the effects of removing acne, treating toxin, whitening skin, clearing eyes, nebula and the like. The pearl powder has wide application in the fields of medicine, material, skin care products and the like, and has medical curative effects of accelerating wound healing and preventing wound inflammation (Hsin-Ling Yang, antibacterial and antioxidant properties of pearl powder 2,2' -azobis (2-amidinopropane) having low-chlorinated-induced homology and oxidative dye to oxidative dye membranes and proteins, journal of Food and Drug Analysis, 2017). The pearl powder is usually further processed by pulverizing, emulsifying and liquefying the lower layer of pearl to prepare drugs, health products, cosmetics and other comprehensive pearl products (Hui-Fang Chiu, efficiency of protein rich pearl powder on antioxidant status in a random decorated tablet-controlled three, Journal of Food and Drug Analysis, 2018). The existing pearl powder hydrolysis technology is divided into water-soluble technology and water-insoluble technology, and the water-soluble technology generally adopts lactic acid hydrolysis method, acid-enzyme hydrolysis method, hydrochloric acid hydrolysis method, ultrasonic hydrolysis method, microwave hydrolysis method, enzyme hydrolysis method, acid hydrolysis method and the like. The water insoluble technology is to process nanometer pearl powder as material through low temperature vacuum drying, supersonic airflow crushing, spray drying and other physical methods. If the Pearl powder is hydrolyzed by neutral protease and lactic acid, the water-soluble Pearl powder is prepared by optimizing the process parameters of enzyme addition, enzymolysis temperature, enzymolysis time and lactic acid dosage by taking the hydrolysis degree as an index, and the molecular mass distribution and the capability of eliminating superoxide anion free radicals of the Pearl powder are measured (Yi-Chen Li, Pearl extract enhancement of the morphology in a surrounding health model, Pharmaceutical Biology, 2013). Or adding bromelain solution and papain solution into nanometer Margarita powder as raw materials, dissolving, vacuum drying at low temperature under reduced pressure, and supersonic jet pulverizing to obtain nanometer Margarita powder rich in active peptide.
The pearl powder hydrolysis technology influences the antioxidation and whitening effects of the pearl extract. "active polypeptide" generally refers to a polypeptide containing 2 to 10 amino acids, the molecular weight is below 2kDa, and it is an active substance, functional factor, with important physiological functions. The pearl extract mainly containing pearl polypeptide is added into B16 melanoma cell culture solution to research the whitening and skin-care effects of the pearl extract. In the aspect of melanin inhibition, the pearl extract can inhibit tyrosinase activity more obviously than arbutin, and the half-inhibition amount of the pearl extract is 80mg/L (poplar is safe, the whitening effect research of the pearl extract, pharmaceutical biotechnology, 2016). Researches show that 0.25%, 0.5% and 0.75% of pearl hydrolysate can inhibit melanin synthesis of melanocytes, and 3 concentrations of pearl hydrolysate can inhibit expression of related gene TRP1 mRNA (Deng Yun, whitening action mechanism research of pearl hydrolysate, Chinese modern traditional medicine, 2017).
Yang safety et al invented a method for separating pearl active polypeptide from pearl protein by enzymolysis, and stated the function of pearl protein in skin whitening and skin care. Adding phosphate buffer solution and neutral protease into Margarita protein solution, and inactivating enzyme in boiling water bath after enzyme catalysis reaction to obtain Margarita active polypeptide (publication No. CN 104911239A).
The invention relates to a method for extracting nano pearl organic matrix and grading molecular weight, which is a method for separating a first pearl organic matrix extract with the molecular weight of more than 5kDa and a second pearl organic matrix extract with the molecular weight of less than 5kDa (publication number CN101381400A) through a centrifugal concentration separation mode of a centrifugal concentration filter membrane device.
Wuwanying et al invented a method for preparing pearl protein, pearl protein prepared by the method and application thereof, adding hydrochloric acid or acetic acid into pearl powder, stirring and centrifuging to obtain precipitate, and freeze-drying the obtained precipitate to obtain the keratin. The protein has good anti-tyrosinase activity, and can be used as raw material of whitening products for medicine and cosmetics (publication number CN 106866807A).
Disclosure of the invention
The invention aims to provide the pearl powder polypeptide extract with the antioxidation effect and the preparation method thereof, the method is convenient, safe and low in cost, the extraction rate of the pearl powder water-soluble protein is effectively improved, and the extract has good hydroxyl free radical removing effect and skin whitening effect.
The technical scheme adopted by the invention is as follows:
a pearl powder polypeptide extract with antioxidant effect is prepared by the following steps:
(1) after the nano pearl powder is dried in vacuum at 40-60 ℃, weighing the pearl powder, adding 2-3 mol/L acetic acid aqueous solution according to the solid-liquid mass ratio of 1: 10-1: 30, magnetically stirring for 2-5 h at 60-80 rpm/min, filtering, and taking supernatant to obtain a crude protein extract; if the solid-liquid ratio is too low, the pearl powder is easy to be insufficiently dissolved; if the concentration of acetic acid is too high, protein inactivation is easily caused; the pH value of the acetic acid ranges from 2.1 to 2.2, the pearl powder is added to react completely, the pH value of the solution ranges from 6.5 to 7.0, and the step of adjusting the pH value by alkali liquor is omitted;
(2) performing rotary evaporation and concentration on the crude protein extract obtained in the step (1) until the volume of the crude protein extract is 1/8-1/10, dialyzing the crude protein extract for 24-36 hours by using a 500Da dialysis bag, and changing water once every 2-3 hours (because the salt concentration in the sample is too high, the volume of the dialyzed sample expands to 3-4 times of the original volume);
(3) and (3) carrying out rotary evaporation and concentration on the dialyzed sample in the step (2) to 1/8-1/10 of the original volume, and carrying out freeze drying to obtain the pearl powder polypeptide extract with the antioxidation effect.
The method of the invention keeps the initial structure of the pearl protein and lays a foundation for analyzing the activity and the structure of the protein in the later period. Compared with the traditional enzymolysis and precipitation treatment method, the method has the advantages that more water-soluble protein components are reserved.
The invention also relates to a method for preparing the pearl powder polypeptide extract with the antioxidation effect, which comprises the following steps:
(1) after the nano pearl powder is dried in vacuum at 40-60 ℃, weighing the pearl powder, adding 2-3 mol/L acetic acid aqueous solution according to the solid-liquid mass ratio of 1: 10-1: 30, magnetically stirring for 2-5 h at 60-80 rpm/min, filtering, and taking supernatant to obtain a crude protein extract;
(2) performing rotary evaporation and concentration on the crude protein extract obtained in the step (1) until the volume of the crude protein extract is 1/8-1/10 of the original volume, dialyzing for 24-36 hours by using a 500Da dialysis bag, and changing water once every 2-3 hours; because the salt concentration in the sample is too high, the volume of the dialysis sample is expanded to 3-4 times of the original volume;
(3) and (3) carrying out rotary evaporation and concentration on the dialyzed sample in the step (2) to 1/8-1/10 of the original volume, and carrying out freeze drying to obtain the pearl powder polypeptide extract with the antioxidation effect.
Preferably, the rotary evaporation in the steps (2) and (3) is carried out at 40-50 ℃ and under the vacuum degree of 0.05-0.1 Mpa. The method has mild reaction conditions (normal temperature of 20-30 ℃), and the temperature of 40-50 ℃ in the concentration process, and fully ensures the enzyme activity. The components are concentrated before final freeze-drying, acetic acid has pungent smell, acetic acid solvent is completely evaporated, partial water is reserved for subsequent freeze-drying, and subsequent application is not influenced.
Specifically, the method comprises the following steps:
(1) after the nano pearl powder is dried in vacuum at 40-60 ℃, weighing the pearl powder, adding 2-3 mol/L acetic acid aqueous solution according to the solid-liquid mass ratio of 1: 10-1: 30, magnetically stirring for 2-5 h at 60-80 rpm/min, filtering by a 0.45 mu m filter membrane, and taking the supernatant to obtain a crude protein extract;
(2) carrying out rotary evaporation concentration on the crude protein extract obtained in the step (1) at the temperature of 40-50 ℃ and the vacuum degree of 0.05-0.1 Mpa until the crude protein extract is 1/10 with the original volume, dialyzing by a 500Da dialysis bag for 24h, and changing water once every 2 h; because the salt concentration in the sample is too high, the volume of the dialysis sample is expanded to 3-4 times of the original volume;
(3) and (3) rotationally evaporating and concentrating the dialyzed sample in the step (2) to 1/10 of the original volume under the conditions of 40-50 ℃ and 0.05-0.1 Mpa of vacuum degree, and freeze-drying to obtain the pearl powder polypeptide extract with the antioxidation effect.
The invention also relates to application of the pearl powder polypeptide extract in preparing antioxidant and whitening cosmetics.
The beneficial effects of the invention are mainly reflected in that: the invention adopts normal-temperature acetic acid solution for auxiliary extraction to obtain a matrix protein extract with high water-soluble protein purity (more than or equal to 40.05 percent), the obtained proteins are all below 10,000Da, the polypeptide content of 0-2000 Da accounts for more than 70 percent of the total protein, the absorption by a human body is facilitated, and the matrix protein extract has obvious hydroxyl free radical removing effect in vitro; and the method has the advantages of simple process, no fine separation step, high preparation efficiency and low cost, and is suitable for subsequent pilot plant test and industrial production.
(IV) description of the drawings
FIG. 1 is a flow chart of the preparation process of the pearl powder polypeptide extract of the invention;
FIG. 2 is a measurement of hydroxyl radical scavenging ability of crude protein of water-soluble pearl powder;
FIG. 3 is a generalized relative chromatogram of water-soluble pearl protein.
(V) detailed description of the preferred embodiments
For the purpose of enhancing understanding of the present invention, the present invention will be described in further detail with reference to specific examples, which are provided for illustration only and are not intended to limit the scope of the present invention:
example 1:
nanometer Margarita powder (obtained by culturing Concha Margaritifera, provided by Zhejiang Changsheng bird Pearl Biotech Co., Ltd.) is dried under vacuum at 50 deg.C (German Binder VD23 vacuum drying oven) → 30g of Margarita powder is weighed, 300g of 3mol/L acetic acid aqueous solution → magnetic stirring for 4h, 60rpm/min (magnetic stirrer, 85-1, Hangzhou David science and education apparatus Co., Ltd.), the liquid is light yellow → filtering (filtering membrane 0.45 μm), supernatant is obtained to obtain water soluble crude protein extract (filter residue is water insoluble crude protein solid) → rotary evaporation (rotary evaporator, R2002B, circulating water pump, Shanghai Shensheng technology Co., Ltd.), 50 deg.C, vacuum degree 0.1Mpa, concentration to original volume 1/10 (about 30mL) → dialysis 24h (500Da dialysis bag), water is changed for 1 time per 2h (volume expansion to 90mL) → rotary evaporation (rotary evaporator), R2002B, circulating water pump, Shanghai Shensheng science and technology Co., Ltd.), 50 deg.C, vacuum degree 0.1Mpa, concentrating to original volume 1/10 to obtain Margarita powder polypeptide extract → measuring water soluble protein content, performing in vitro hydroxyl free radical scavenging activity evaluation → lyophilizing to obtain Margarita powder polypeptide extract (water soluble protein), and performing GPC component analysis on its components.
Through analysis and detection, the maximum in-vitro hydroxyl free radical clearance rate of the pearl powder polypeptide extract can reach 77.88 percent, and the IC of the pearl powder polypeptide extract is50The yield was 1.78% and the protein purity was 41.90% when the protein was 190.80 μ g/mL.
Evaluation of in vitro hydroxyl radical scavenging effect activity:
0.5mL of a 6mmol/L salicylic acid-absolute ethanol solution, 0.5mL of a 9mmol/L ferrous sulfate solution and 0.5mL of a 0.072% hydrogen peroxide solution were sequentially added to 0.5mL of a sample solution (the sample was a freeze-dried solid powder dissolved in distilled water at concentrations of 100, 150, 200, 250 and 300mg/mL respectively), and distilled water was added to make the total volume 2.5mL, and the mixture was subjected to a water bath at 37 ℃ for 30min, cooled in cold water, and then the absorbance was measured at 510 nm. VCAs a control. The formula of the free radical clearance rate is as follows:
clearance (%) ═ a0-(Ai-Aj))/A0x100％
(A0: 0.5mL of 95% ethanol solution (blank absorbance); a. thei: 0.5mL sample (sample absorbance); a. thej: 0.5mL sample +0.5mL 95% ethanol solution (sample background absorbance)
Half maximal Inhibitory Concentration (IC) of free radical scavenging activity50) The values are represented.
And (3) GPC analysis and detection:
yield of extract: (dry weight of extract/dry weight of pearl powder raw material) x 100%
② content of water-soluble protein: and (3) determining by using a BCA protein quantitative kit method.
③ GPC analysis: GPC System (waters 1525)&Agilent PL-GPC220, USA). Mobile phase: 0.2M NaNO3 0.01M NaH2PO4. A chromatographic column: PL aquqgel-OH MIXED 8 μm. Flow rate 1.0mL min-1The amount of the sample was 10.0. mu.L. Column temperature: at 30 ℃. Operating time: and 15 min. The initial mobile phase was used to equilibrate for 30min between each injection. The proportion of the molecular weight of each segment is calculated by a molecular weight relative distribution table, and the result is as follows:
table 1: the yield of each component of pearl powder protein
As can be seen from Table 1, the yield of the crude protein extract of pearl powder is 2.3-2.5% (the protein content of pearl powder is about 3%, the yield is that the finally obtained protein accounts for the mass percent of the pearl powder), the yield of the water-soluble protein component (the polypeptide extract of pearl powder) is 1.5-1.8%, and the yield of the water-insoluble protein component is 0.5-1.0%. The measurement result of the hydroxyl radical scavenging ability of crude protein of water-soluble pearl powder is shown in FIG. 2, and the IC is calculated50The value is 124-125 mg/mL.
Table 2: generalized relative peak table of water-soluble pearl protein
The generalized relative chromatogram of the water-soluble pearl protein is shown in figure 3, and the generalized relative peak value table of the water-soluble pearl protein is shown in table 2; as can be seen, the number average molecular weight of the water-soluble pearl protein is 904Da, the weight average molecular weight is 1735Da, the dispersity is about 1.92, the relative molecular mass of the protein is small, and most of the protein is small molecular peptide.
Table 3: molecular weight relative distribution table of water-soluble pearl protein
The purity of the protein in the sample is about 41.9 percent, the table 3 is the ratio data of the molecular weight of the water-soluble protein in the sample, the content of 5000-7000 Da is 0.62-0.75 percent, and the content of the molecular weight is lower. The content of 1000-2000 Da is 43.51-47.26%, and the molecular weight of the molecular weight is highest. The content of 0-1000 Da is 27.53-29.03%, and the content of 2000-5000 Da is 24.32-28.05%.
Example 2:
nanometer Margarita powder (Zhejiang Changsheng Pearl Biotechnology Co., Ltd.) is dried under vacuum at 40 deg.C (German Binder VD23 vacuum drying oven) → 30g of Margarita powder is weighed, 600g of 4mol/L acetic acid water solution → magnetic stirring for 2h, 80rpm/min (magnetic stirrer, 85-1, Hangzhou David science and education apparatus Co., Ltd.), the liquid is orange yellow, filtering (filtering membrane 0.45 μm), supernatant is obtained to obtain crude protein extract → rotary evaporation (rotary evaporator, R2002B, circulating water pump, Shanghai Shensheng technology Co., Ltd.), 48 deg.C, vacuum degree 0.1Mpa, concentrating to original volume 1/10 (about 60mL) → dialyzing 24h (500Da dialysis bag), water is changed for 1 time every 2h (volume amplification is 240mL) → rotary evaporation (rotary evaporator, R2002B, circulating water pump, Shanghai Shensheng Sheng Co., Ltd.), concentrating at 48 deg.C under 0.1Mpa to original volume of 1/10 → measuring water soluble protein content, performing in vitro hydroxyl free radical scavenging activity evaluation → lyophilizing to obtain Margarita powder polypeptide extract, and performing GPC component analysis. Through analysis and detection, the maximum in-vitro hydroxyl free radical clearance rate of the pearl powder polypeptide extract can reach 75.25 percent, and the IC of the pearl powder polypeptide extract is50The yield was 1.63% and the protein purity was 40.05% when the protein was 190.16 μ g/mL.
Example 3:
nanometer Margarita powder (Zhejiang Changsheng bird pearl Biotechnology Co., Ltd.) is dried under vacuum at 40 deg.C (German Binder VD23 vacuum drying oven) → 30g of Margarita powder is weighed, 600g of 2mol/L acetic acid water solution → magnetic stirring is carried out for 5h, 60rpm/min (magnetic stirrer, 85-1, Hangzhou David scientific and education apparatus Co., Ltd.) is added, the liquid is light yellow → filtering (filtering membrane 0.45 μm), and supernatant is taken to obtain crude protein extract → rotary evaporation (rotary evaporation)An evaporator, R2002B, a circulating water pump, Shanghai Shensheng science and technology limited), 47 ℃, the vacuum degree of 0.1Mpa, the concentration to the original volume of 1/10 (about 60mL) → dialysis for 24h (500Da dialysis bag), water exchange for 1 time every 2h → rotary evaporation (rotary evaporator, R2002B, a circulating water pump, Shanghai Shensheng science and technology limited), 47 ℃, the vacuum degree of 0.1Mpa, the concentration to the original volume of 1/10 (volume expansion is 210mL) → measurement of the content of water-soluble protein, in-vitro hydroxyl radical scavenging activity evaluation → freeze drying is carried out, and the polypeptide pearl powder extract is obtained and subjected to GPC component analysis. Through analysis and detection, the maximum in-vitro hydroxyl free radical clearance rate of the pearl powder polypeptide extract can reach 80.76 percent, and the IC of the pearl powder polypeptide extract is50The yield was 1.80% and the protein purity was 45.61% at 185.85 μ g/mL.
Claims (5)
1. A pearl powder polypeptide extract with antioxidant effect is prepared by the following steps:
(1) after the nano pearl powder is dried in vacuum at 40-60 ℃, weighing the pearl powder, adding 2-3 mol/L acetic acid aqueous solution according to the solid-liquid mass ratio of 1: 10-1: 30, magnetically stirring for 2-5 h at 60-80 rpm/min, filtering, and taking supernatant to obtain a crude protein extract;
(2) performing rotary evaporation and concentration on the crude protein extract obtained in the step (1) until the volume of the crude protein extract is 1/8-1/10 of the original volume, dialyzing for 24-36 hours by using a 500Da dialysis bag, and changing water once every 2-3 hours;
(3) and (3) carrying out rotary evaporation and concentration on the dialyzed sample in the step (2) to 1/8-1/10 of the original volume, and carrying out freeze drying to obtain the pearl powder polypeptide extract with the antioxidation effect.
2. A method for preparing the pearl powder polypeptide extract having an antioxidant effect of claim 1, which comprises:
(1) after the nano pearl powder is dried in vacuum at 40-60 ℃, weighing the pearl powder, adding 2-3 mol/L acetic acid aqueous solution according to the solid-liquid mass ratio of 1: 10-1: 30, magnetically stirring for 2-5 h at 60-80 rpm/min, filtering, and taking supernatant to obtain a crude protein extract;
(2) performing rotary evaporation and concentration on the crude protein extract obtained in the step (1) until the volume of the crude protein extract is 1/8-1/10 of the original volume, dialyzing for 24-36 hours by using a 500Da dialysis bag, and changing water once every 2 hours;
(3) and (3) carrying out rotary evaporation and concentration on the dialyzed sample in the step (2) to 1/8-1/10 of the original volume, and carrying out freeze drying to obtain the pearl powder polypeptide extract with the antioxidation effect.
3. The method of claim 2, wherein the rotary evaporation in steps (2) and (3) is carried out at 40 to 50 ℃ and a vacuum of 0.05 to 0.1 MPa.
4. The method of claim 2, characterized in that the method is as follows:
(1) after the nano pearl powder is dried in vacuum at 40-60 ℃, weighing the pearl powder, adding 2-3 mol/L acetic acid aqueous solution according to the solid-liquid mass ratio of 1: 10-1: 30, magnetically stirring for 2-5 h at 60-80 rpm/min, filtering by a 0.45 mu m filter membrane, and taking the supernatant to obtain a crude protein extract;
(2) carrying out rotary evaporation concentration on the crude protein extract obtained in the step (1) at the temperature of 40-50 ℃ and the vacuum degree of 0.05-0.1 Mpa until the crude protein extract is 1/10 with the original volume, dialyzing by a 500Da dialysis bag for 24h, and changing water once every 2 h;
(3) and (3) rotationally evaporating and concentrating the dialyzed sample in the step (2) to 1/10 of the original volume under the conditions of 40-50 ℃ and 0.05-0.1 Mpa of vacuum degree, and freeze-drying to obtain the pearl powder polypeptide extract with the antioxidation effect.
5. The use of the pearl powder polypeptide extract of claim 1 in the preparation of anti-oxidant and whitening cosmetics.
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| CN115991758A (en) * | 2022-08-27 | 2023-04-21 | 广西中医药大学 | Pearl powder polypeptide for promoting diabetic wound healing and application thereof |
| WO2024000089A1 (en) * | 2022-06-27 | 2024-01-04 | 欧诗漫生物股份有限公司 | Method for preparing pearl-whitening and anti-aging combined polypeptide and single whitening polypeptide and use thereof |
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